MiR-138 and MiR-135 directly target focal adhesion kinase, inhibit cell invasion, and increase sensitivity to chemotherapy in cancer cells.

Abstract

Focal Adhesion Kinase is a 125 kDa non-receptor kinase and overexpressed in many types of tumors. Recently, short noncoding RNAs, called microRNAs have been discovered as regulators of gene expression mainly through binding to the untranslated region (UTR) of mRNA. In this report we show that MiR-138 and MiR-135 down-regulated FAK expression in cancer cells. MiR-138 and MiR-135 inhibited FAK protein expression in different cancer cell lines. The computer analysis of 3'FAK-untranslated region (FAKUTR) identified one conserved MiR-138 binding site (CACCAGCA) at positions 3514-3521 and one conserved MiR-135 (AAGCCAU) binding site at positions 4278-4284 in the FAK-UTR. By a dual-luciferase assay we demonstrate that MiR-138 and MiR-135 directly bound the FAK untranslated region using FAK-UTR-Target (FAK-UTR) luciferase plasmid and inhibited its luciferase activity. The sitedirected mutagenesis of the MiR-138 and MiR-135 binding sites in the FAK-UTR abrogated MiR-138 and MiR-135-directed inhibition of FAK-UTR. Real-time PCR demonstrated that cells transfected with MiR-138 and MiR-135 expressed decreased FAK mRNA levels. Moreover, stable expression of MiR-138 and MiR-135 in 293 and HeLa cells decreased cell invasion and increased sensitivity to 5- fluorouracil (5-FU), FAK inhibitor, Y15, and doxorubicin. In addition, MiR-138 significantly decreased 293 xenograft tumor growth in vivo. This is the first report on regulation of FAK expression by MiR-135 and MiR138 that affected invasion, drug sensitivity, and tumor growth in cancer cells, which is important to the development of FAK-targeted therapeutics and understanding their novel regulations and functions.

• Authors: Vita M Golubovskaya, Brittany Sumbler, Baotran Ho, Michael Yemma, William G Cance
• Journal: Anti-cancer agents in medicinal chemistry (Anticancer Agents Med Chem) Vol. 14 Issue 1 Pg. 18-28 (Jan 2014) ISSN: 1875-5992 [Electronic] Netherlands
• PMID: 23438844 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• 1,2,4,5-benzenetetraamine
• 3' Untranslated Regions
• Aniline Compounds
• Antineoplastic Agents
• MIRN135 microRNA, human
• MIRN138 microRNA, human
• MicroRNAs
• RNA, Messenger
• Doxorubicin
• Focal Adhesion Protein-Tyrosine Kinases
• Fluorouracil

Topics

• 3' Untranslated Regions
• Aniline Compounds (pharmacology)
• Animals
• Antineoplastic Agents (pharmacology)
• Doxorubicin (pharmacology)
• Female
• Fluorouracil (pharmacology)
• Focal Adhesion Protein-Tyrosine Kinases (antagonists & inhibitors, genetics, metabolism)
• Gene Expression Regulation, Neoplastic (drug effects, genetics)
• HEK293 Cells
• HeLa Cells
• Heterografts
• Humans
• Mice
• Mice, Nude
• MicroRNAs (genetics, metabolism)
• Neoplasm Invasiveness (genetics, pathology)
• RNA, Messenger (metabolism)